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S. Jasty, S. Krishnakumar, P. Gunisha, N. Archana, G. Suganeswari, L. Gopal; Retinal Stem/Progenitor Properties of Ciliary Epithelial and Iris pigment Epithelial Cells From the Adult Human Cadaveric Eye. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5030. doi: https://doi.org/.
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Stem/progenitor cells that give rise to neurons and glia have been identified in several regions of the brain including the embryonic retina and the ciliary epithelium of the adult mice, chick, pigs etc., raising the possibility of autologous transplantation. The presence of progenitor cells in various regions of the adult eye includes the Ciliary body and Iris regions, which are readily accessible during routine eye surgery. We studied the presence of retinal stem/progenitors using the cadaveric human eye.
Ciliary Epithelium (CE) and Iris Pigment Epithelium (IPE) cells from cadaveric humans were cultured in the presence of mitogens like Epidermal growth factor (EGF) and Fibroblast growth factor (FGF) to generate neurospheres. Seven days after seeding the growth was evaluated under phase contrast microscope and the expression of several markers for the stem/progenitor cells was done by RT-PCR and Immunofluorescence studies. The generated neurospheres were plated on laminin-coated slides in the presence of the Brain derived neurotrophic factor (BDNF) and Retinoic acid for the neurosphere differentiation.
An average of 0.5-1.0 x 104 cells/ml were isolated from each of the tissue. The cells isolated from CE and IPE proliferated to form Neurospheres. These Neurospheres were formed in serum-free medium and were positive for ABCG2 (Universal stem cell markers) and Nestin, Beta tubulin III, Musashi1 (early neuronal marker) and PAX6 (neuro-epithelial progenitor markers). Neurospheres were also BrdU positive indicating that the cells are proliferating. The differentiating markers like the Retinal ganglion cells (Brn3b, Thy-1, Calretinin) and Photoreceptor cells (s-opsin, Rhodopsin, Recoverin) were expressed.
This study suggests that ciliary epithelium (CE) and Iris pigment epithelium (IPE) in the adult human eye harbours a mitotic quiescent population of neural stem cells and can be grown in vitro, thereby providing potential sources of donor cells for transplantation. The CE and IPE retinal stem/progenitors proliferate in the presence of mitogens and share the expression of universal neural and retinal progenitor markers. The analysis of the differentiation potential of CE and IPE stem cells shows that they are capable of generating both early (e.g., retinal ganglion cells) and late (e.g., rod photoreceptors) born retinal neurons.
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